*Target (protein/gene name): Leishmania major, acidocalcisomal exopolyphosphatase *NCBI Gene # or RefSeq#:12983123 *Protein ID (NP or XP #) or Wolbachia#: LmjF01.0310 *Organism (including strain):Leishmania major strain Friedlin Etiologic Risk Group (see link below):Risk group 2 (RG2) - parasitic agents *Background/Disease Information: Leishmania major is a protozoa associated with the disease zoonotic cutaneous, also known as (also known as "Aleppo boil," "Baghdad boil," "Bay sore," "Biskra button," "Chiclero ulcer," "Delhi boil," "Kandahar sore," "Lahore sore," "Oriental sore," "Pian bois," and "Uta"). This disease is the acute form of cutaneous leishmaniasis, and it affects people living in rural areas, specifically Northern Africa, the Middle East, Northwestern China and Northwestern India, near infected rats. Symptoms include skin lesions that spread rapidly and become severely inflamed. There is no known treatment to date. Link to TDR Targets page (if present): http://www.tdrtargets.org/targets/view?gene_id=29931 Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): http://www.ncbi.nlm.nih.gov/gene/?term=LmjF_01_0310 Essentiality of this protein:
LmjF01.0310 has essentiality data
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in bloodstream forms (3 days); Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in bloodstream forms (6 days); Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in procyclic forms; Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in differentiation of procyclic to bloodstream forms; Complex of proteins?: This target does not have a complex of proteins. Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): As of now, there are no chemical compounds associated with this gene.
Structure Available (PDB or Homology model)
-- PDB #: 3RF0
Expression Information (has it been expressed in bacterial cells): Yes, it has been expressed in E. coli. Purification Method: Affinity chromatography is the purification method used. Image of protein (PyMol with features delineated and shown separately):
Figure 2 - Exopolyphosphatase from Yersinia pestis (3RF0), protein shown as cartoon, colored from the N-terminus to the C-terminus in rainbow
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*length of your protein in Amino Acids: 519 amino acids Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 58711.2 Da Molar Extinction coefficient of your protein at 280 nm wavelength: 56630 M-1 cm-1 TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*GC% Content for gene: 59.7% *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): **
*NCBI Gene # or RefSeq#: 12983123
*Protein ID (NP or XP #) or Wolbachia#: LmjF01.0310
*Organism (including strain): Leishmania major strain Friedlin
Etiologic Risk Group (see link below): Risk group 2 (RG2) - parasitic agents
*Background/Disease Information:
Leishmania major is a protozoa associated with the disease zoonotic cutaneous, also known as (also known as "Aleppo boil," "Baghdad boil," "Bay sore," "Biskra button," "Chiclero ulcer," "Delhi boil," "Kandahar sore," "Lahore sore," "Oriental sore," "Pian bois," and "Uta"). This disease is the acute form of cutaneous leishmaniasis, and it affects people living in rural areas, specifically Northern Africa, the Middle East, Northwestern China and Northwestern India, near infected rats. Symptoms include skin lesions that spread rapidly and become severely inflamed. There is no known treatment to date.
Link to TDR Targets page (if present):
http://www.tdrtargets.org/targets/view?gene_id=29931
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):
http://www.ncbi.nlm.nih.gov/gene/?term=LmjF_01_0310
Essentiality of this protein:
LmjF01.0310 has essentiality data
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in bloodstream forms (3 days); Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in bloodstream forms (6 days); Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in procyclic forms; Source study: alsford
Gene/Ortholog: Tb09.160.1950 (OG4_13037); Phenotype: no significant loss or gain of fitness in differentiation of procyclic to bloodstream forms;
Complex of proteins?: This target does not have a complex of proteins.
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): As of now, there are no chemical compounds associated with this gene.
*EC#: 3.6.1.11
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/php/result_flat.php4?ecno=3.6.1.11
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
http://www.sigmaaldrich.com/catalog/papers/20633993
-- links to assay reagents (substrates) pages.
http://www.ncbi.nlm.nih.gov/pubmed/?term=acidocalcisomal+exopolyphosphatase
Structure Available (PDB or Homology model)
-- PDB #: 3RF0
Expression Information (has it been expressed in bacterial cells): Yes, it has been expressed in E. coli.
Purification Method: Affinity chromatography is the purification method used.
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*length of your protein in Amino Acids: 519 amino acids
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 58711.2 Da
Molar Extinction coefficient of your protein at 280 nm wavelength: 56630 M-1 cm-1
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
*GC% Content for gene: 59.7%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**